anti cd40  (Sino Biological)


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    Sino Biological anti cd40
    Anti Cd40, supplied by Sino Biological, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cd40/product/Sino Biological
    Average 91 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    anti cd40 - by Bioz Stars, 2022-10
    91/100 stars

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    Sino Biological anti cd40l monoclonal abs
    Targeting wild-type or codon-optimized HA2 via <t>CD40L</t> provides complete heterosubtypic protection. Survival curves and body weight loss of Balb/c mice immunized with two doses of 10 9 pfu of the indicated rAd constructs and challenged with 5 × LD 50 of mouse-adapted influenza ( a ) A/Puerto Rico/8/1934(H1N1) ( b ) A/Jingke/30/95(H3N2) or ( c ) A/chicken/Jiangsu/7/2002(H9N2) viruses 2 weeks post secondary immunization. Data are shown from one experiment with n =10 mice per treatment group.
    Anti Cd40l Monoclonal Abs, supplied by Sino Biological, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cd40l monoclonal abs/product/Sino Biological
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cd40l monoclonal abs - by Bioz Stars, 2022-10
    91/100 stars
      Buy from Supplier

    91
    Sino Biological mouse cd40
    The superior rigidity of IgG2 CH1-hinge. a A diagram showing anti-mCD40 antibody TR-FRET. Anti-mCD40 antibody molecules mixed with <t>CD40-Tb</t> and CD40-D2 (mCD40 labeled with Tb donor and D2 acceptor fluorochromes, respectively) can simultaneously bind CD40-Tb and CD40-D2, and emit TR-FRET signal quantified as the relative ratio of detected 665 nm fluorescence to 620 nm fluorescence (Em665/Em620) upon stimulation. b A diagram of the model showing that hinge flexibility of hIgG anti-mCD40 antibodies correlates with TR-FRET signal levels. Left, anti-mCD40 antibodies with little hinge flexibility do not trigger TR-FRET signal due to the large distance between CD40-Tb and CD40-D2; middle, hinge flexibility can bring CD40-Tb and CD40-D2 close enough to trigger TR-FRET signal; right, anti-mCD40 antibodies with large hinge flexibility give rise to stronger TR-FRET signal due to more molecules with closer CD40 binding sites. c – e TR-FRET signal levels of anti-mCD40 antibodies of indicated constant domains. Shown are relative TR-FRET signal levels (Em665/Em620) plotted against the concentration of control IgG or the anti-mCD40 antibodies of indicated constant domains. Bars represent the mean ± SEM. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001, two-way ANOVA with Holm–Sidak’s post hoc. Source data ( c – e ) are provided as a Source Data file. A representative of two independent experiments is shown
    Mouse Cd40, supplied by Sino Biological, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse cd40/product/Sino Biological
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse cd40 - by Bioz Stars, 2022-10
    91/100 stars
      Buy from Supplier

    88
    Sino Biological antibody drug
    The superior rigidity of IgG2 CH1-hinge. a A diagram showing anti-mCD40 antibody TR-FRET. Anti-mCD40 antibody molecules mixed with <t>CD40-Tb</t> and CD40-D2 (mCD40 labeled with Tb donor and D2 acceptor fluorochromes, respectively) can simultaneously bind CD40-Tb and CD40-D2, and emit TR-FRET signal quantified as the relative ratio of detected 665 nm fluorescence to 620 nm fluorescence (Em665/Em620) upon stimulation. b A diagram of the model showing that hinge flexibility of hIgG anti-mCD40 antibodies correlates with TR-FRET signal levels. Left, anti-mCD40 antibodies with little hinge flexibility do not trigger TR-FRET signal due to the large distance between CD40-Tb and CD40-D2; middle, hinge flexibility can bring CD40-Tb and CD40-D2 close enough to trigger TR-FRET signal; right, anti-mCD40 antibodies with large hinge flexibility give rise to stronger TR-FRET signal due to more molecules with closer CD40 binding sites. c – e TR-FRET signal levels of anti-mCD40 antibodies of indicated constant domains. Shown are relative TR-FRET signal levels (Em665/Em620) plotted against the concentration of control IgG or the anti-mCD40 antibodies of indicated constant domains. Bars represent the mean ± SEM. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001, two-way ANOVA with Holm–Sidak’s post hoc. Source data ( c – e ) are provided as a Source Data file. A representative of two independent experiments is shown
    Antibody Drug, supplied by Sino Biological, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/antibody drug/product/Sino Biological
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    antibody drug - by Bioz Stars, 2022-10
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    Targeting wild-type or codon-optimized HA2 via CD40L provides complete heterosubtypic protection. Survival curves and body weight loss of Balb/c mice immunized with two doses of 10 9 pfu of the indicated rAd constructs and challenged with 5 × LD 50 of mouse-adapted influenza ( a ) A/Puerto Rico/8/1934(H1N1) ( b ) A/Jingke/30/95(H3N2) or ( c ) A/chicken/Jiangsu/7/2002(H9N2) viruses 2 weeks post secondary immunization. Data are shown from one experiment with n =10 mice per treatment group.

    Journal: Mucosal Immunology

    Article Title: Targeting the HA2 subunit of influenza A virus hemagglutinin via CD40L provides universal protection against diverse subtypes

    doi: 10.1038/mi.2014.59

    Figure Lengend Snippet: Targeting wild-type or codon-optimized HA2 via CD40L provides complete heterosubtypic protection. Survival curves and body weight loss of Balb/c mice immunized with two doses of 10 9 pfu of the indicated rAd constructs and challenged with 5 × LD 50 of mouse-adapted influenza ( a ) A/Puerto Rico/8/1934(H1N1) ( b ) A/Jingke/30/95(H3N2) or ( c ) A/chicken/Jiangsu/7/2002(H9N2) viruses 2 weeks post secondary immunization. Data are shown from one experiment with n =10 mice per treatment group.

    Article Snippet: Forty-eight hours later, protein expression in cell lysates of infected cells was confirmed by western blot of proteins separated on sodium dodecyl sulfate -polyacrylamide gel electrophoresis or native gradient (4–12%) polyacrylamide gel electrophoresis as previously described using anti-CD40L monoclonal Abs and rabbit anti-HA Abs (Sino Biological Inc., Beijing, China).

    Techniques: Mouse Assay, Construct

    CD40L and HA2 codon-optimization enhance circulating and nasal anti-HA2 antibody response. HA2-specific antibody titers at 2 weeks after ( a ) priming and ( b ) boosting are shown for circulating IgG1 and IgG2a, and nasal IgA. Balb/c mice were intranasally immunized with 10 9 pfu of each rAd virus or with PBS on day 0 and boosted 2 weeks post primary immunization. Data are shown as mean titer±s.d. from one experiment out of three independent experiments, with n =3 mice per treatment group in each experiment. **** P

    Journal: Mucosal Immunology

    Article Title: Targeting the HA2 subunit of influenza A virus hemagglutinin via CD40L provides universal protection against diverse subtypes

    doi: 10.1038/mi.2014.59

    Figure Lengend Snippet: CD40L and HA2 codon-optimization enhance circulating and nasal anti-HA2 antibody response. HA2-specific antibody titers at 2 weeks after ( a ) priming and ( b ) boosting are shown for circulating IgG1 and IgG2a, and nasal IgA. Balb/c mice were intranasally immunized with 10 9 pfu of each rAd virus or with PBS on day 0 and boosted 2 weeks post primary immunization. Data are shown as mean titer±s.d. from one experiment out of three independent experiments, with n =3 mice per treatment group in each experiment. **** P

    Article Snippet: Forty-eight hours later, protein expression in cell lysates of infected cells was confirmed by western blot of proteins separated on sodium dodecyl sulfate -polyacrylamide gel electrophoresis or native gradient (4–12%) polyacrylamide gel electrophoresis as previously described using anti-CD40L monoclonal Abs and rabbit anti-HA Abs (Sino Biological Inc., Beijing, China).

    Techniques: Mouse Assay

    Enhanced protection and immune response is dependent on CD40L. Survival curves and body weight loss of Balb/c mice immunized with two doses of 10 9 pfu of the indicated rAd constructs and challenged with 5 × LD 50 of mouse-adapted influenza ( a ) A/Puerto Rico/8/1934(H1N1) ( b ) A/Jingke/30/95(H3N2) or ( c ) A/chicken/Jiangsu/7/2002(H9N2) viruses 2 weeks post secondary immunization. Circulating HA2-specific IgG1 and IgG2a titers ( d ) and IFN-γ and IL-4T cell responses ( e ) are shown at 2 weeks post secondary immunization. Data are shown from one experiment with n =8 mice per treatment group in ( a–c ). Data in ( d ) are shown as mean titer±s.d. from one experiment, with n =5 mice per treatment group. Data in ( e ) are shown as mean numbers of spot-forming cells (SFCs)±s.d. from one experiment with n =5. Each sample was tested in triplicates. **** P

    Journal: Mucosal Immunology

    Article Title: Targeting the HA2 subunit of influenza A virus hemagglutinin via CD40L provides universal protection against diverse subtypes

    doi: 10.1038/mi.2014.59

    Figure Lengend Snippet: Enhanced protection and immune response is dependent on CD40L. Survival curves and body weight loss of Balb/c mice immunized with two doses of 10 9 pfu of the indicated rAd constructs and challenged with 5 × LD 50 of mouse-adapted influenza ( a ) A/Puerto Rico/8/1934(H1N1) ( b ) A/Jingke/30/95(H3N2) or ( c ) A/chicken/Jiangsu/7/2002(H9N2) viruses 2 weeks post secondary immunization. Circulating HA2-specific IgG1 and IgG2a titers ( d ) and IFN-γ and IL-4T cell responses ( e ) are shown at 2 weeks post secondary immunization. Data are shown from one experiment with n =8 mice per treatment group in ( a–c ). Data in ( d ) are shown as mean titer±s.d. from one experiment, with n =5 mice per treatment group. Data in ( e ) are shown as mean numbers of spot-forming cells (SFCs)±s.d. from one experiment with n =5. Each sample was tested in triplicates. **** P

    Article Snippet: Forty-eight hours later, protein expression in cell lysates of infected cells was confirmed by western blot of proteins separated on sodium dodecyl sulfate -polyacrylamide gel electrophoresis or native gradient (4–12%) polyacrylamide gel electrophoresis as previously described using anti-CD40L monoclonal Abs and rabbit anti-HA Abs (Sino Biological Inc., Beijing, China).

    Techniques: Mouse Assay, Construct

    CD40L enhances T-cell responses to HA2. Balb/c mice were intranasally immunized twice with 10 9 pfu of each rAd virus or with PBS. Mice were sacrificed at 2 weeks ( a ) post primary or ( b ) secondary immunization and splenocytes were isolated for IFN-γ, TNF-α, and IL-4 measurement by ELISpot. Data shown as mean numbers of spot-forming cells (SFCs)±s.d. from one experiment with n =3 per treatment group. Each sample was tested in triplicates. **** P

    Journal: Mucosal Immunology

    Article Title: Targeting the HA2 subunit of influenza A virus hemagglutinin via CD40L provides universal protection against diverse subtypes

    doi: 10.1038/mi.2014.59

    Figure Lengend Snippet: CD40L enhances T-cell responses to HA2. Balb/c mice were intranasally immunized twice with 10 9 pfu of each rAd virus or with PBS. Mice were sacrificed at 2 weeks ( a ) post primary or ( b ) secondary immunization and splenocytes were isolated for IFN-γ, TNF-α, and IL-4 measurement by ELISpot. Data shown as mean numbers of spot-forming cells (SFCs)±s.d. from one experiment with n =3 per treatment group. Each sample was tested in triplicates. **** P

    Article Snippet: Forty-eight hours later, protein expression in cell lysates of infected cells was confirmed by western blot of proteins separated on sodium dodecyl sulfate -polyacrylamide gel electrophoresis or native gradient (4–12%) polyacrylamide gel electrophoresis as previously described using anti-CD40L monoclonal Abs and rabbit anti-HA Abs (Sino Biological Inc., Beijing, China).

    Techniques: Mouse Assay, Isolation, Enzyme-linked Immunospot

    Recombinant Ad constructs and in vitro protein expression. ( a ) Schematic representation of the generated rAd constructs. The rAd-SHA2(Opt)FCD40L and rAd-SHA2(wt)FCD40L were generated to express secreted HA2(Opt)-FCD40L and HA2(wt)-FCD40L fusion proteins, respectively. S is an N-terminal leader sequence derived from human tyrosinase signal peptide. HA2 is the HA2 subunit from influenza A/California/7/2009 virus(H1N1). F is a 27 amino-acid fragment from the T4 bacteriophage fibritin trimerization motif fused with ectodomain (144 amino acids) of the mouse CD40L. The rAd-SHA2(Opt)F expresses secreted trimeric HA2 protein. The rAd-SHA2(wt) and rAd-SHA2(Opt) express secreted monomeric HA2(wt) and HA2(Opt), respectively. The rAd-control is an “empty” control vector. Constructs were engineered in recombinant ΔE1/E3 adenovirus pAdxsi vectors under the control of CMV promoter. ( b ) In vitro protein expression in cell culture. Confluent HeLa cells were infected with rAds at multiplicity of infection of 100 and 48 h later, cell lysates were collected. Protein expression was confirmed by western blot using anti-HA2 polyclonal antibodies.

    Journal: Mucosal Immunology

    Article Title: Targeting the HA2 subunit of influenza A virus hemagglutinin via CD40L provides universal protection against diverse subtypes

    doi: 10.1038/mi.2014.59

    Figure Lengend Snippet: Recombinant Ad constructs and in vitro protein expression. ( a ) Schematic representation of the generated rAd constructs. The rAd-SHA2(Opt)FCD40L and rAd-SHA2(wt)FCD40L were generated to express secreted HA2(Opt)-FCD40L and HA2(wt)-FCD40L fusion proteins, respectively. S is an N-terminal leader sequence derived from human tyrosinase signal peptide. HA2 is the HA2 subunit from influenza A/California/7/2009 virus(H1N1). F is a 27 amino-acid fragment from the T4 bacteriophage fibritin trimerization motif fused with ectodomain (144 amino acids) of the mouse CD40L. The rAd-SHA2(Opt)F expresses secreted trimeric HA2 protein. The rAd-SHA2(wt) and rAd-SHA2(Opt) express secreted monomeric HA2(wt) and HA2(Opt), respectively. The rAd-control is an “empty” control vector. Constructs were engineered in recombinant ΔE1/E3 adenovirus pAdxsi vectors under the control of CMV promoter. ( b ) In vitro protein expression in cell culture. Confluent HeLa cells were infected with rAds at multiplicity of infection of 100 and 48 h later, cell lysates were collected. Protein expression was confirmed by western blot using anti-HA2 polyclonal antibodies.

    Article Snippet: Forty-eight hours later, protein expression in cell lysates of infected cells was confirmed by western blot of proteins separated on sodium dodecyl sulfate -polyacrylamide gel electrophoresis or native gradient (4–12%) polyacrylamide gel electrophoresis as previously described using anti-CD40L monoclonal Abs and rabbit anti-HA Abs (Sino Biological Inc., Beijing, China).

    Techniques: Recombinant, Construct, In Vitro, Expressing, Generated, Sequencing, Derivative Assay, Plasmid Preparation, Cell Culture, Infection, Western Blot

    The superior rigidity of IgG2 CH1-hinge. a A diagram showing anti-mCD40 antibody TR-FRET. Anti-mCD40 antibody molecules mixed with CD40-Tb and CD40-D2 (mCD40 labeled with Tb donor and D2 acceptor fluorochromes, respectively) can simultaneously bind CD40-Tb and CD40-D2, and emit TR-FRET signal quantified as the relative ratio of detected 665 nm fluorescence to 620 nm fluorescence (Em665/Em620) upon stimulation. b A diagram of the model showing that hinge flexibility of hIgG anti-mCD40 antibodies correlates with TR-FRET signal levels. Left, anti-mCD40 antibodies with little hinge flexibility do not trigger TR-FRET signal due to the large distance between CD40-Tb and CD40-D2; middle, hinge flexibility can bring CD40-Tb and CD40-D2 close enough to trigger TR-FRET signal; right, anti-mCD40 antibodies with large hinge flexibility give rise to stronger TR-FRET signal due to more molecules with closer CD40 binding sites. c – e TR-FRET signal levels of anti-mCD40 antibodies of indicated constant domains. Shown are relative TR-FRET signal levels (Em665/Em620) plotted against the concentration of control IgG or the anti-mCD40 antibodies of indicated constant domains. Bars represent the mean ± SEM. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001, two-way ANOVA with Holm–Sidak’s post hoc. Source data ( c – e ) are provided as a Source Data file. A representative of two independent experiments is shown

    Journal: Nature Communications

    Article Title: Human immunoglobulin G hinge regulates agonistic anti-CD40 immunostimulatory and antitumour activities through biophysical flexibility

    doi: 10.1038/s41467-019-12097-6

    Figure Lengend Snippet: The superior rigidity of IgG2 CH1-hinge. a A diagram showing anti-mCD40 antibody TR-FRET. Anti-mCD40 antibody molecules mixed with CD40-Tb and CD40-D2 (mCD40 labeled with Tb donor and D2 acceptor fluorochromes, respectively) can simultaneously bind CD40-Tb and CD40-D2, and emit TR-FRET signal quantified as the relative ratio of detected 665 nm fluorescence to 620 nm fluorescence (Em665/Em620) upon stimulation. b A diagram of the model showing that hinge flexibility of hIgG anti-mCD40 antibodies correlates with TR-FRET signal levels. Left, anti-mCD40 antibodies with little hinge flexibility do not trigger TR-FRET signal due to the large distance between CD40-Tb and CD40-D2; middle, hinge flexibility can bring CD40-Tb and CD40-D2 close enough to trigger TR-FRET signal; right, anti-mCD40 antibodies with large hinge flexibility give rise to stronger TR-FRET signal due to more molecules with closer CD40 binding sites. c – e TR-FRET signal levels of anti-mCD40 antibodies of indicated constant domains. Shown are relative TR-FRET signal levels (Em665/Em620) plotted against the concentration of control IgG or the anti-mCD40 antibodies of indicated constant domains. Bars represent the mean ± SEM. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001, two-way ANOVA with Holm–Sidak’s post hoc. Source data ( c – e ) are provided as a Source Data file. A representative of two independent experiments is shown

    Article Snippet: CD40-binding ELISA and FACS Binding of anti-CD40 antibodies to mouse CD40 was analyzed by ELISA.

    Techniques: Labeling, Fluorescence, Binding Assay, Concentration Assay

    Divergent hIgG agonism and specific FcγR-binding requirement. a Diagram showing the OVA-specific CD8 + T-cell response model. In brief, FcγR-humanized (hFCGR Tg ) or -deficient (FcγRα −/− ) mice were adoptively transferred with OT-I cells on day-1, immunized intraperitoneally (i.p.) with 2 μg of DEC-OVA in the presence of control or anti-CD40 antibodies on day 0. Splenocytes were harvested to quantify OVA-specific CD8 + T cells on day 6. b , c Representative FACS profile ( b ) and quantification ( c ) showing the percentage of OT-I cells (CD45.1 + TCRVα2 + ) among CD8 + T cells in mice treated and analyzed as in ( a ) together with 30 μg of indicated control or anti-mCD40 antibodies. Numbers of mice: b , c three hFCGR Tg mice for Ctrl IgG, five hFCGR Tg , and three FcγRα −/− mice for other groups. d – f Quantification of OT-I cells as the percentage of OT-I cells among CD8 + T cells ( d , e ) or cell count ( f ) in FcγR-humanized mice treated and analyzed as in ( a ) together with 10 μg ( d , e ) or 30 μg ( f ) of indicated control or anti-mCD40 antibodies (the N297A mutation abrogates Fc–FcγR binding) and with/without FcγRIIB-blocking antibody 2B6 (150 μg per mouse) ( f ). Numbers of mice: d four mice per group; e five mice per group; f two mice for Ctrl IgG, five mice for αmCD40:G2, six mice for αmCD40:G2 + 2B6. g Quantification of OT-I cells as the percentage of OT-I cells among CD8 + T cells in mice of indicated genotypes (FcγR-deficient (FcgRα −/− , five mice per group), FcγRIIB-deficient (Fcgr2b −/− , five mice per group) or humanized (Fcgr2b −/− hFCGR2B Tg , four mice per group)) treated and analyzed as in ( a ) together with 10 μg of indicated control or IgG2 anti-mCD40 antibodies. Each symbol represents an individual mouse. Bars represent the mean ± SEM. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001; unpaired two-tailed t test ( c , g ), one-way ANOVA with Holm–Sidak’s post hoc ( d – f ). Source data ( c – g ) are provided as a Source Data file. A representative of two independent experiments is shown

    Journal: Nature Communications

    Article Title: Human immunoglobulin G hinge regulates agonistic anti-CD40 immunostimulatory and antitumour activities through biophysical flexibility

    doi: 10.1038/s41467-019-12097-6

    Figure Lengend Snippet: Divergent hIgG agonism and specific FcγR-binding requirement. a Diagram showing the OVA-specific CD8 + T-cell response model. In brief, FcγR-humanized (hFCGR Tg ) or -deficient (FcγRα −/− ) mice were adoptively transferred with OT-I cells on day-1, immunized intraperitoneally (i.p.) with 2 μg of DEC-OVA in the presence of control or anti-CD40 antibodies on day 0. Splenocytes were harvested to quantify OVA-specific CD8 + T cells on day 6. b , c Representative FACS profile ( b ) and quantification ( c ) showing the percentage of OT-I cells (CD45.1 + TCRVα2 + ) among CD8 + T cells in mice treated and analyzed as in ( a ) together with 30 μg of indicated control or anti-mCD40 antibodies. Numbers of mice: b , c three hFCGR Tg mice for Ctrl IgG, five hFCGR Tg , and three FcγRα −/− mice for other groups. d – f Quantification of OT-I cells as the percentage of OT-I cells among CD8 + T cells ( d , e ) or cell count ( f ) in FcγR-humanized mice treated and analyzed as in ( a ) together with 10 μg ( d , e ) or 30 μg ( f ) of indicated control or anti-mCD40 antibodies (the N297A mutation abrogates Fc–FcγR binding) and with/without FcγRIIB-blocking antibody 2B6 (150 μg per mouse) ( f ). Numbers of mice: d four mice per group; e five mice per group; f two mice for Ctrl IgG, five mice for αmCD40:G2, six mice for αmCD40:G2 + 2B6. g Quantification of OT-I cells as the percentage of OT-I cells among CD8 + T cells in mice of indicated genotypes (FcγR-deficient (FcgRα −/− , five mice per group), FcγRIIB-deficient (Fcgr2b −/− , five mice per group) or humanized (Fcgr2b −/− hFCGR2B Tg , four mice per group)) treated and analyzed as in ( a ) together with 10 μg of indicated control or IgG2 anti-mCD40 antibodies. Each symbol represents an individual mouse. Bars represent the mean ± SEM. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001; unpaired two-tailed t test ( c , g ), one-way ANOVA with Holm–Sidak’s post hoc ( d – f ). Source data ( c – g ) are provided as a Source Data file. A representative of two independent experiments is shown

    Article Snippet: CD40-binding ELISA and FACS Binding of anti-CD40 antibodies to mouse CD40 was analyzed by ELISA.

    Techniques: Binding Assay, Mouse Assay, FACS, Cell Counting, Mutagenesis, Blocking Assay, Two Tailed Test

    CH1-hinge basis of agonistically inactive IgG3 and superior IgG2. a – c Quantification of OT-I cells as the percentage of OT-I cells among CD8 + T cells ( a ) or cell count ( b , c ) in FcγR-humanized mice treated and analyzed as in Fig. 1a together with indicated control or anti-mCD40 antibodies ( a 3.16 μg per mouse; b , c 10 μg per mouse). Numbers of mice: a six mice for Ctrl IgG, six to seven mice per group for others; b , c three mice for Ctrl IgG, four to five mice per group for others. d – f Representative FACS profile ( d ) and quantification ( e , f ) showing the percentage of OT-I cells (CD45.1 + TCRVα2 + ) among CD8 + T cells in human CD40/FcγR-transgenic mice treated and analyzed as in Fig. 1a together with control or anti-human CD40 antibodies of indicated clones (Clones 21.4.1 and 3.1.1 have been described in Patent No.:US 7,338,660; Clone 21.4.1 in the IgG2 form is also known as CP-870,893) and constant domains (30 μg per mouse). Numbers of mice: d – f five to six mice per group. Each symbol represents an individual mouse. Bars represent the mean ± SEM. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001; one-way ANOVA with Holm–Sidak’s post hoc. Source data ( a – c , e , f ) are provided as a Source Data file. A representative of two independent experiments is shown

    Journal: Nature Communications

    Article Title: Human immunoglobulin G hinge regulates agonistic anti-CD40 immunostimulatory and antitumour activities through biophysical flexibility

    doi: 10.1038/s41467-019-12097-6

    Figure Lengend Snippet: CH1-hinge basis of agonistically inactive IgG3 and superior IgG2. a – c Quantification of OT-I cells as the percentage of OT-I cells among CD8 + T cells ( a ) or cell count ( b , c ) in FcγR-humanized mice treated and analyzed as in Fig. 1a together with indicated control or anti-mCD40 antibodies ( a 3.16 μg per mouse; b , c 10 μg per mouse). Numbers of mice: a six mice for Ctrl IgG, six to seven mice per group for others; b , c three mice for Ctrl IgG, four to five mice per group for others. d – f Representative FACS profile ( d ) and quantification ( e , f ) showing the percentage of OT-I cells (CD45.1 + TCRVα2 + ) among CD8 + T cells in human CD40/FcγR-transgenic mice treated and analyzed as in Fig. 1a together with control or anti-human CD40 antibodies of indicated clones (Clones 21.4.1 and 3.1.1 have been described in Patent No.:US 7,338,660; Clone 21.4.1 in the IgG2 form is also known as CP-870,893) and constant domains (30 μg per mouse). Numbers of mice: d – f five to six mice per group. Each symbol represents an individual mouse. Bars represent the mean ± SEM. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001; one-way ANOVA with Holm–Sidak’s post hoc. Source data ( a – c , e , f ) are provided as a Source Data file. A representative of two independent experiments is shown

    Article Snippet: CD40-binding ELISA and FACS Binding of anti-CD40 antibodies to mouse CD40 was analyzed by ELISA.

    Techniques: Cell Counting, Mouse Assay, FACS, Transgenic Assay, Clone Assay

    Impact of both IgG CH1-hinge and Fc on anti-CD40 antibody antitumour activities. MC38 ( a , b ) and MO4 ( c ) tumour volumes in FcγR-humanized mice following treatment with control or anti-mCD40 antibodies of indicated constant domains. After tumour cells were subcutaneously inoculated and established in FcγR-humanized mice, mice were treated i.p. twice on day 0 (the day when mice with palpable tumours receive their first treatment) and day 3 with 31.6 μg/mouse of control or anti-CD40 antibodies of indicated constant domains (the N297A mutation abrogates Fc–FcγR binding), and monitored for tumour growth. For mice inoculated with MO4 tumour cells in ( c ), each treatment also included 2 μg/mouse of DEC-OVA. Shown are tumour growth curves of individual mice ( a ) or mouse groups ( b , c ). Numbers of mice: a , b seven to eight mice per group; c seven mice per group except six mice for αmCD40:G2(N297A). Bars represent the mean ± SEM. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001, chi-square test ( a ), and two-way ANOVA with Holm–Sidak’s post hoc ( b , c ) were used for group comparison. Source data ( a – c ) are provided as a Source Data file. A representative of two independent experiments is shown ( a , b )

    Journal: Nature Communications

    Article Title: Human immunoglobulin G hinge regulates agonistic anti-CD40 immunostimulatory and antitumour activities through biophysical flexibility

    doi: 10.1038/s41467-019-12097-6

    Figure Lengend Snippet: Impact of both IgG CH1-hinge and Fc on anti-CD40 antibody antitumour activities. MC38 ( a , b ) and MO4 ( c ) tumour volumes in FcγR-humanized mice following treatment with control or anti-mCD40 antibodies of indicated constant domains. After tumour cells were subcutaneously inoculated and established in FcγR-humanized mice, mice were treated i.p. twice on day 0 (the day when mice with palpable tumours receive their first treatment) and day 3 with 31.6 μg/mouse of control or anti-CD40 antibodies of indicated constant domains (the N297A mutation abrogates Fc–FcγR binding), and monitored for tumour growth. For mice inoculated with MO4 tumour cells in ( c ), each treatment also included 2 μg/mouse of DEC-OVA. Shown are tumour growth curves of individual mice ( a ) or mouse groups ( b , c ). Numbers of mice: a , b seven to eight mice per group; c seven mice per group except six mice for αmCD40:G2(N297A). Bars represent the mean ± SEM. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001, chi-square test ( a ), and two-way ANOVA with Holm–Sidak’s post hoc ( b , c ) were used for group comparison. Source data ( a – c ) are provided as a Source Data file. A representative of two independent experiments is shown ( a , b )

    Article Snippet: CD40-binding ELISA and FACS Binding of anti-CD40 antibodies to mouse CD40 was analyzed by ELISA.

    Techniques: Mouse Assay, Mutagenesis, Binding Assay